An image capture system and method for imaging biological samples disposed in culture media supported by a plate. The system has a calibration module, an image acquisition module and an image presentation module. When the system receives a culture plate for imaging, default values for the culture plate and media are used to begin image acquisition at a given time. The captured image is then used to create a pixel by pixel map of the image. The system inspects the pixel-by-pixel map for saturated pixels and for signal to noise ratio and acquires a new image if the number of saturated pixels is at or above a predetermined threshold or the signal to noise ratio for the pixel is below a predetermined threshold. From this inspection a new value of photon flux and/or exposure time is determined and a new image is captured using the new value and the steps are repeated. Upon a determination that a predetermined threshold signal to noise ratio has been obtained for the non-saturated pixels, or when the predetermined upper threshold for the time interval for image acquisition is elapsed the system provides a final image for the given time.
Legal claims defining the scope of protection, as filed with the USPTO.
1. A system for imaging biological samples disposed in culture media, the system comprising: a system calibration module that provides default values for capturing an image of a biological sample disposed on culture media disposed in a plate; an image acquisition module comprising a camera wherein the image acquisition module is adapted to acquire data for a series of images at a given time interval, in communication with the system calibration module, the image acquisition configured to: i) acquire data for a first image using default values for photon flux and exposure time from the system calibration module and creating a pixel by pixel map of the image data, each pixel associated with a signal to noise ratio, a photon flux and exposure time, and an intensity; ii) update the image acquisition by reviewing the image data to identify saturated pixels and selecting one of a new photon flux, a new exposure time or both based on whether a ratio of saturated to non-saturated pixels is greater than or less than a predetermined saturation threshold and, based on that determination; iii) use a new value for the photon flux, the exposure time or both to acquire data for a new image and iv) update the map of the image data with the new values for signal to noise ratio, the photon flux, the exposure time or both and pixel intensity, wherein the image acquisition module is configured to acquire data for a new image if the signal to noise ratio of unsaturated pixels is less than a predetermined SNR threshold or a number of saturated pixels exceeds the predetermined saturation threshold, wherein the image acquisition module finalizes image acquisition when either the signal to noise ratio of unsaturated pixels meets or exceeds the predetermined SNR threshold, a predetermined allocated time for image acquisition has elapsed or a predetermined maximum number of images has been acquired; and an image presentation module that transforms the image data from the image acquisition module into an image for viewing or analysis.
2. The system of claim 1 wherein the photon flux is a set value and the system controls a camera sensor integration by controlling exposure time wherein the calibration module determines default photon flux and exposure time for a variety of plates and culture media.
3. The system of claim 2 wherein the new value for the photon flux is obtained by using either a new exposure time or a new light intensity value or both.
4. The system of claim 1 wherein signal to noise ratio is determined for at least a portion of the image of the sample disposed on the culture media.
5. The system of claim 1 wherein the image acquisition module acquires image data from the camera for at least one or more channels or one or more spectral bands.
6. The system of claim 1 wherein the image acquisition module assigns a grey value for each pixel for each image acquisition, and the grey value for each pixel is updated after each image acquisition.
7. The system of claim 6 wherein the updated grey value is the previous grey value minus a predetermined reference value, wherein the predetermined reference value is a predetermined value based on the plate, the plate media and the exposure time for the image acquisition module assigns a grey value for each pixel for each acquisition, and the grey value for each pixel is updated after each image acquisition.
8. The system of claim 1 wherein the image acquisition module is configured to operate in at least one of an automatic mode where all pixels are treated equally or in a supervised mode, where the analyzed pixels are those having been identified as associated with one or more objects of interest.
9. The system of claim 1 wherein the photon flux is a set value and the system controls a camera sensor integration by controlling exposure time.
10. A method for imaging biological samples disposed in culture media, the method comprising: determining default values for obtaining an image of a biological sample disposed on culture media supported in a plate; acquiring image data corresponding to a series of images at a first time over a first time interval, the data of the first image in the series being acquired using predetermined defaults for photon flux and exposure time; creating a pixel by pixel map of the image data; associating data for each pixel with a signal to noise ratio, a value for photon flux, exposure time, and an intensity; updating at least one of the image photon flux value and exposure time value by: i) reviewing the image data for saturated pixels and the signal to noise ratio for the pixels and selecting a new value for at least one of photon flux and exposure time based on whether a ratio of saturated to non-saturated pixels is greater than or less than a predetermined threshold and whether the signal to noise ratio of unsaturated pixels meets or exceeds a predetermined SNR threshold, and based on that determination, using the new value for photon flux, exposure time, or both to acquire a new image and updating the map of the image with the new values for signal to noise ratio, photon flux and exposure time, and pixel intensity; ii) acquiring data for the new image using the at least one new value for photon flux and exposure time; iii) optionally repeating steps i) and ii); finalizing image data acquisition for the time interval when either the image data is at or above the predetermined SNR threshold, a predetermined maximum allotted time for image acquisition has elapsed or a predetermined maximum number of images has been acquired; and repeating the steps of acquiring, creating, associating, updating and finalizing for a second time over a second time interval; and transforming the image data acquired at first and second time intervals into first and second images the first image acquired at the first time and the second image acquired at a second time.
11. The method of claim 10 wherein the value for photon flux is constant and the exposure time value updated and wherein the predetermined default values are based on plate, plate culture and exposure time.
12. The method of claim 10 further comprising determining the pixels for which the image map will be created, wherein the pixels are associated with an object of interest.
13. The method of claim 12 wherein the default values comprise a black level for the pixels associated with an object of interest at a default exposure time.
14. The method of claim 10 wherein the pixel by pixel map is a grey value, the signal to noise ratio and the exposure time for each pixel.
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January 30, 2015
February 5, 2019
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